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2006
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In the Laboratory
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Preparation, Purification, and Secondary Structure Determination of Bacillus circulans Xylanase. A Modular Laboratory Incorporating Aspects of Molecular Biology, Biochemistry, and Biophysical Chemistry
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Sal Russo and Lisa Gentile
Department of Chemistry, Western Washington University, Bellingham, WA 98225-9150
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December 2006
Vol. 83 No. 12
p. 1850
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| Abstract |
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An integrated six-week module that allows students to develop biochemistry laboratory techniques in the context of a discovery-oriented project has been developed. This module, suitable for undergraduate third- or fourth-year biochemistry lab, focuses on protein over-expression, purification, and stability of Bacillus circulans xylanase (BCX). BCX is an ideal protein to study as it has important industrial applications and is highly over-expressed, easily purified in a single chromatographic step, and stable. The discovery-based aspect of the project monitors, by circular dichroism spectroscopy, the secondary structure of BCX under conditions that compromise its stabilizing intramolecular forces. Results are written up as a formal lab report in the format of an article in Biochemistry. This module aids in student learning for several reasons: (i) by engaging students in multiple techniques in the context of a single protein, they understand how each technique contributes to the overall production and characterization of BCX, (ii) by allowing students to design a CD-based module, they are required to explore more deeply the forces responsible for protein stability, and (iii) by combining molecular biology, biochemistry, and biophysical chemistry, students more easily see connections between the disciplines.
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| Supplement |
Instructor notes, experimental procedures, and student handouts are available.
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| More Information |
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Citation |
Russo, Sal; Gentile, Lisa. J. Chem. Educ. 2006 83 1850.
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Keywords |
Biochemistry; Biophysical Chemistry; Enzymes; Hands-On Learning / Manipulatives; Inquiry-Based / Discovery Learning; Interdisciplinary / Multidisciplinary; Ion Exchange; Laboratory Instruction; Molecular Properties / Structure; Proteins / Peptides; Spectroscopy; Upper-Division Undergraduate
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History |
Created:
Last Updated: |
10/30/2006
11/7/2006
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 Caution! | | Experiments, laboratory exercises, lecture demonstrations, and other descriptions of the use of chemicals, apparatus, instruments, computers, and computer interfaces are presented in the Journal of Chemical Education as illustrative of new or improved ideas or concepts in chemistry instruction and are directed at qualified teachers. Although every effort is made to assure and encourage safe practices and safe use of chemicals, the Journal of Chemical Education cannot assume responsibility for uses made of its published materials. Many chemicals are hazardous. Precautions for the safe use of hazardous chemicals and directions for their proper disposal are described in the Material Safety Data Sheets and on the labels. We strongly urge all those planning to use materials from our pages to make choices and to develop procedures for laboratory and classroom safety in accordance with local needs and situations. |
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